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Method for selectively producing lignin-degrading enzymes by using phanerochaete chrysosporium

A technology for lignin-degrading enzyme and Phoenicia protothecoides, which is applied in the biological field and can solve the problems that the selective production of lignin-degrading enzymes of P. protochaetans is not seen, the effect is not ideal, and the like.

Inactive Publication Date: 2015-01-07
TSINGHUA UNIV
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  • Abstract
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Problems solved by technology

Li Huazhong et al. (2002) also studied Phanerochaete chrysosporium on Mn 2+ Selective production of lignin-degrading enzymes under concentration adjustment, but the effect is not ideal and is still limited by pure oxygen supply
[0005] Except Mn 2+ Control. At present, there are no reports on the selective production of Phanerochaete chrysosporium lignin-degrading enzymes using other regulation methods, especially in the air environment

Method used

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  • Method for selectively producing lignin-degrading enzymes by using phanerochaete chrysosporium
  • Method for selectively producing lignin-degrading enzymes by using phanerochaete chrysosporium
  • Method for selectively producing lignin-degrading enzymes by using phanerochaete chrysosporium

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Experimental program
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Effect test

Embodiment 1

[0033] Embodiment 1, Phanerochaete chrysosporium selectively produces lignin peroxidase under carbon limitation

[0034] 1. Preparation of carbon-limited liquid medium

[0035] A carbon-limited liquid medium was used, and the medium was slightly modified based on the typical Tien and Kirk medium (1988). The C / N ratio (molar ratio) of the medium was 3.8, and the specific composition was as follows:

[0036] The carbon-limited liquid medium consists of a final concentration of 5.04g / L (28mM) glucose, a final concentration of 4.05g / L (NH 4 + 44mM) ammonium tartrate, the final concentration is 2.0g / L KH 2 PO 4 , the final concentration is 0.5g / L MgSO 4 , the final concentration is 0.1g / L CaCl 2 , the final concentration is 1mg / L vitamin B 1 , final concentration of 1.5mM veratrol, final concentration of 20mM (pH 4.4) acetate buffer, 70mL / L trace element solution and deionized water; pH is 4.4.

[0037] The trace element solution was prepared with a final concentration of 3g...

Embodiment 2

[0051] Embodiment 2, Phanerochaete chrysosporium selectively produces manganese peroxidase under nitrogen limitation

[0052] 1. Preparation of nitrogen-limited liquid medium

[0053] Nitrogen-limited liquid medium is adopted, and the C / N ratio (molar ratio) of the medium is 152.7, and the specific composition is as follows:

[0054] Nitrogen-limited liquid medium formula is the final concentration of 10.08g / L (56mM) glucose, 0.203g / L (NH 4 + 2.2mM) ammonium tartrate replaces glucose and ammonium tartrate in the carbon-limited liquid medium, and the remaining components are the same as the carbon-limited liquid medium in Example 1; pH is 4.4.

[0055] 2. Selective production of manganese peroxidase by Phanerochaete chrysosporium under nitrogen limitation

[0056] The culture conditions and analysis methods are the same as in Example 1, except that the culture medium is replaced by a nitrogen-limited liquid medium.

[0057] Lignin-degrading enzymes produce results such as ...

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Abstract

The invention relates to a method for selectively producing lignin-degrading enzymes by using phanerochaete chrysosporium. The invention provides a method for selectively producing lignin-degrading enzymes by using phanerochaete chrysosporium, which comprises the following steps: fermenting phanerochaete chrysosporium in a culture medium containing an immobilized culture carrier in a non-immersed state, and collecting a fermentation product, namely, a lignin-degrading enzyme. Experiments show that when the method is adopted for carrying out fermentation on a lignin-degrading enzyme, the lignin-degrading enzyme can be selectively obtained under air conditions without feeding pure oxygen or air containing high-concentration oxygen in the process of culturing but just by controlling the concentration of a carbon and nitrogen source in the culture medium and adding a carrier in a non-immersed state. The method disclosed by the invention is simple in condition control, and the obtained lignin-degrading enzyme is strong in selectivity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for selectively producing lignin degrading enzymes by using Phanerochaete chrysosporium. Background technique [0002] White rot fungus is a kind of higher filamentous fungus, most of which belong to Basidiomycetes in classification, grow on trees or wood, and get its name because it causes woody white rot. White-rot fungi degrade lignin because they can produce lignin-degrading enzymes and secrete them outside the cells. The degradation of lignin by lignin-degrading enzymes is a chain reaction process based on free radicals, which has strong substrate non-specificity and oxidation. This degradation mechanism enables lignin-degrading enzymes to not only degrade lignin, but also degrade Many xenobiotics and persistent toxic organic pollutants in the environment (Barr & Aust, 1994; Cameron et al., 2000; Gao et al., 2010). Therefore, white-rot fungi and their lignin-degrading...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/08C12R1/645
CPCC12N9/0065C12Y111/01013C12Y111/01014
Inventor 文湘华喻国策钱易王建龙
Owner TSINGHUA UNIV
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